Segmental Adeno-Cre Infection: A Technique to Generate Isolated Colorectal Cancer using Genetically Engineered Mouse Models
Published: April 30, 2023
Abstract
Source: Betzler, A. M., et al. A Genetically Engineered Mouse Model of Sporadic Colorectal Cancer. J. Vis. Exp. (2017).
This video describes the generation of a colorectal mouse model by segmental adeno-cre infection. The infected colon segment leads to tumorigenesis within this segment of the mucosa, resulting in adenomas ultimately progressing to invasive and metastatic carcinoma. This model is an attractive platform for studies of cancer biology and preclinical therapeutic trials.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Local Tumor Induction via Surgical Adeno-cre Infection Preparation of animals for surgery NOTE: Virtually any conditional ("floxed") mutation can be induced via the here described method. The use of mutations in genes relevant to colorectal canc…
Representative Results
Figure 1. Colonoscopic Images of Colorectal Tumors (Conditional Alleles of the Given Animal in Brackets). A. Normal distal colon. B. Early adenoma 2 weeks after adeno-cre infection. Note the green color due to a GFP reporter allele in this mouse. (Apctm2Rak, Krastm4Tyj, Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze<…
開示
The authors have nothing to disclose.
Materials
| Dulbecco's Phosphate Buffered Saline | Life Technologies GmbH | 14190169 | |
| Trypsin-EDTA (0.25%, PhenolRed) | Life Technologies GmbH | 25200072 | |
| Normal saline 0.9% (E154) | Serumwerk Bernburg AG | 10013 | |
| Aqua ad injectabilia | B. Braun Melsungen AG | 235144 | |
| Ad5CMV-Cre (adenovirus, c = 2E +11 PFU/mL) | Gene Transfer Vector Core University of Iowa | ||
| 15 mL, 50 mL centrifuge tubes | Greiner Bio-One GmbH | 188271/227270 | |
| Eppendorf tubes 1.5 mL/ 2 mL | Sarstedt AG & Co. | 72,695,400 | |
| Petri dish PS 100/15 mm (sterile, Nuclon) | Fisher Scientific GmbH | 10508921/ NUNC150350 | |
| 1 mL Syringe (without dead volume) – Injekt-F SOLO | Braun/neoLab | 194291661 | |
| 30G injection needle | BECTON DICKINSON | 304000 | |
| Sevoflurane (Sevoflurane AbbVie) | AbbVie Germany GmbH & Co. KG | ||
| Medical oxygen | Air Liquide Medical GmbH | ||
| Buprenorphine (Temgesic) | Indivior Eu Ltd. | ||
| Bepanthen – ophthalmic ointment | Bayer Vital GmbH | 10047757 | |
| Table Top Research Anesthesia Machine x/O2 Flush w/ Sevoflurane Vaporizer | Parkland Scientific | V3000PS/PK | |
| Cellulose swabs | Lohmann & Rauscher Deutschland | 13356 | |
| Insulin syringe EMG 1 mL (with 30G cannula) | B. Braun Melsungen AG | 9161627S | |
| Fine Bore Tubing (bore: 0.28 mm/ diameter: 0.61mm) | Smiths Medical Deutschland 8 | 800/100/100 | |
| Micro-Adson Forceps | Fine Science Tools | 11018-12 | |
| Iris Scissor – ToughCut | Fine Science Tools | 14058-11 | |
| Olsen-Hegar Needle Holder | Fine Science Tools | 12002-12 | |
| AutoClip Kit | Fine Science Tools | 12020-00 | |
| PDS Z1012H 6/0 C1 (surgical suture) | Johnson & Johnson Medical GmbH | Z1012H | |
| Curved Micro Serrefine Vascular Clamp | Fine Science Tools | 18055-05 | |
| Fogarty Spring Clips | Edwards | CDSAFE 6 | |
| Hot Plate 062 | Labotect | 13854 | |
| Isis – Hair shaver | Aesculap – Braun | ||
| Cold Light Fountain XENON 175 | Karl Storz | 20132101-1 | Karl Storz Coloview System Mainz |
| Fiber Optic Light Cable | Karl Storz | 69495NL | Karl Storz Coloview System Mainz |
| TRICAM Three-Chip Camera Head | Karl Storz | 20221030 | Karl Storz Coloview System Mainz |
| TRICAM SLII Camera Control Unit | Karl Storz | 20223011-1 | Karl Storz Coloview System Mainz |
| 15" Flat Screen Monitor EndoVue | Karl Storz | 9415NN | Karl Storz Coloview System Mainz |
| HOPKINS Straight Forward Telescope diameter 1.9 mm; length 10 cm autoclavable fiber optic light transmission incorporated | Karl Storz | 64301AA | |
| Protection and Examination Sheath | Karl Storz | 61029C |
